KR20220106393A - Cosmetic composition comprising extract of Glycyrrhiza uralensis by-product - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising an extract of Glycyrrhiza uralensis by-products. The cosmetic composition of the present invention exhibits excellent anti-inflammatory, antioxidant and antibacterial activities, contains a high content of polyphenolic compounds and flavonoid compounds, and provides excellent functionality such as prevention of skin trouble caused by inflammation or microorganisms and anti-aging of the skin. In addition, the cosmetic composition of the present invention utilizes by-products of Glycyrrhiza uralensis, which are mostly discarded, so that it is expected to be of great help in alleviating environmental pollution problems caused by the disposal of by-products and providing opportunities for additional income generation to farmers.

Description

Cosmetic composition comprising extract of Glycyrrhiza uralensis by-product

The present invention relates to a cosmetic composition comprising an extract of licorice by-product.

Licorice is added to other medicines in oriental medicine and used as a neutralizing agent (adjusting agent). used to do In addition, glycyrrhizin, the main medicinal component of licorice, is known to lower blood cholesterol levels and lower blood pressure, promote bile secretion, and have antitussive and analgesic effects.

Until now, licorice mainly uses the root part, so most of the other parts, namely, the leaves and stems of licorice, which are by-products that account for more than 50% of the licorice cultivation area, are thrown away. Therefore, it is necessary to develop a technology that utilizes licorice by-products to solve the problem of environmental pollution caused by these discarded by-products and also to generate income for growers.

Accordingly, the present inventors tried to develop a method that can effectively utilize the by-product of licorice, and in particular, studied the possibility of applying it to cosmetics. In the past, a technology for applying licorice to cosmetics has been developed, but since this is a technology using the part used as a herbal medicine, that is, the root part, it was intended to develop a cosmetic that can exhibit excellent functionality by utilizing the discarded licorice by-products.

Korean Patent Registration No. 10-0154366

Therefore, the main object of the present invention is to provide a cosmetic composition that can exhibit excellent functionality using the by-product of licorice, which is mostly thrown away.

According to one aspect of the present invention, the present invention provides a cosmetic composition comprising an extract of licorice by-product.

In the composition of the present invention, it is preferable that the licorice by-product is a leaf of licorice, a licorice, or a mixture thereof.

In the composition of the present invention, the licorice by-product is preferably a mixture of licorice leaves and creepers in a weight ratio of 1: 2 to 2: 1.

In the composition of the present invention, the extract is preferably extracted with water, ethanol or a mixed solution thereof.

In the composition of the present invention, the extract is preferably extracted with 50 to 95% (v/v) ethanol.

In the composition of the present invention, the extract is preferably extracted with 70 to 200 ml of 50 to 95% (v/v) ethanol per 10 g of the licorice by-product.

In the composition of the present invention, the cosmetic is preferably a cosmetic for skin antioxidant, antibacterial or anti-inflammatory.

The cosmetic composition of the present invention exhibits excellent anti-inflammatory, antioxidant and antibacterial activity, and contains a high content of polyphenol compounds and flavonoid compounds to provide excellent functionality such as preventing skin troubles caused by inflammation or microorganisms and preventing skin aging. can In addition, since the cosmetic composition of the present invention utilizes the by-products of licorice, which are mostly discarded, it is expected that it will be of great help in improving the environmental pollution problem caused by the disposal of by-products and providing opportunities for additional income generation to farmers.

1 shows the cytotoxicity test results of licorice by-product extract according to an embodiment of the present invention.
Figure 2 shows the anti-inflammatory test results of licorice by-product extract according to an embodiment of the present invention.
Figure 3 shows the results of the antioxidant experiment of licorice by-product extract according to an embodiment of the present invention.
Figure 4 shows the test result of the total polyphenol content of licorice by-product extract according to an embodiment of the present invention.
5 shows the experimental results of total flavonoid content of licorice by-product extract according to an embodiment of the present invention.

The cosmetic composition of the present invention is characterized in that it comprises an extract of licorice by-product.

The scientific name of licorice is ' Glycyrrhiza uralensis Fischer', and as is well known, its root is mainly used as a herbal medicine.

Licorice in the present invention is preferably domestically grown licorice.

In the present invention, licorice by-product means a part other than the root of licorice, preferably leaves, stems, or mixtures thereof, and more preferably leaves, licorice, or mixtures thereof.

In the case of applying the mixture of the leaves and licorice as a by-product of licorice, preferably licorice leaves and creepers are mixed in a weight ratio of 1: 2 to 2: 1, and more preferably, licorice leaves and creepers are 1: 1.8 to 1.8: mixed in a weight ratio of 1, more preferably, licorice leaves and creeping diameter are mixed in a weight ratio of 1: 1.5 to 1.5: 1, and more preferably, licorice leaves and creeping diameter are 1: 1.3 to 1.3. : mixed in a weight ratio of 1: More preferably, the leaves and creeping diameter of licorice are mixed in a weight ratio of 1: 1.2 to 1.2: 1, more preferably, the leaves and creeping diameter of licorice are 1: 1.1 to 1.1: 1 mixed in a weight ratio of

In the present invention, the extract may be an extract obtained by extracting the juice of licorice by-product or licorice by-product with a solvent. In the case of a juice extract, it can be obtained using a conventional juicer, for example, a juicer used to extract juice from plant materials, and in the case of a solvent extract, it is obtained using a conventional extraction method using a solvent or an extraction apparatus. can do.

As the extraction solvent, various solvents known in the art may be used. The solvent usable in the present invention may be water, an alcohol having 1 to 4 carbon atoms, acetone, hexane, ethyl acetate, carbon dioxide, or a solvent in which two or more thereof are mixed. Preferably, water, ethanol, or a mixed solution thereof is used as the solvent. In order to increase the effect presented in the present invention, preferably 50 to 95% (v/v) ethanol solution, for example, a solution in which water and ethanol are mixed in a volume ratio of 50:50 to 5:95 is used, and more preferably 60 to 90% (v/v), more preferably 70 to 90% (v/v), more preferably 75 to 85% (v/v) ethanol solution is used.

The licorice by-product used as an extraction raw material in the present invention can be applied in various forms, but is preferably applied in the form of a powder. Licorice by-product powder can be obtained through a conventional method of pulverizing plants. At this time, preferably the size of 10 mesh (size that can pass through a sieve of 10 mesh), more preferably the size of 30 mesh (size that can pass through a sieve of 30 mesh), even more preferably 45 It is used after powdering to the size of the mesh (a size that can pass through a sieve of 45 mesh). In order to increase the effect presented in the present invention, it is preferably applied in the form of a powder obtained by performing freeze-drying and then pulverizing, that is, a freeze-dried powder. Freeze-drying of licorice by-products can be obtained through a conventional method of freeze-drying plants.

In the present invention, the extraction method may use various methods known in the art, for example, a method of adding an extraction solvent to an extraction raw material and performing stationary, shaking or ultrasonic extraction. At this time, preferably, 50 to 1000 ml of extraction solvent is added per 10 g of licorice by-product, for example, licorice by-product, which is an extraction raw material, powder or freeze-dried powder, and more preferably 50 to 500 ml of extraction per 10 g of licorice by-product. A solvent is added, and more preferably, 70 to 200 ml of an extraction solvent per 10 g of licorice by-product is added. In addition, the extraction conditions at this time are preferably 1 to 100 hours at 40 to 80 ℃, more preferably 12 to 48 hours at 50 to 70 ℃, more preferably 20 to 30 hours at 55 to 65 ℃. A method of repeating extraction several times, i.e., primary extraction, then filtration to obtain an extract, adding an extraction solvent again to the extraction residue, and repeating the secondary extraction method, and collecting the extract obtained in each order can be used. have. At this time, the repeated extraction will be appropriate for the second to fourth repeated extraction.

After extraction using the solvent as described above, the extract is preferably filtered, more preferably filtered and concentrated under reduced pressure, and more preferably after filtration and concentration under reduced pressure, freeze-drying is used.

According to the present invention, the licorice by-product extract as described above exhibits anti-inflammatory, antioxidant and antibacterial activity, and contains a polyphenol compound and a flavonoid compound, and may be particularly useful in cosmetics applied to the skin.

According to the physiological activity and ingredients contained as described above, the cosmetic composition of the present invention is preferably a cosmetic for skin antioxidant, antibacterial or anti-inflammatory.

According to the experimental results, all of the extracts of Cressaceae, leaf extracts, and mixed extracts of Crested cypress and leaves exhibited excellent antioxidant, antibacterial and anti-inflammatory activities, and exhibited excellent total polyphenol content and total flavonoid content, but among them, leaf extract and specific extracts It was found that the mixed extract of the raw material/extraction solvent ratio was superior in the overall aspect. In consideration of this point, the cosmetic composition of the present invention preferably includes a licorice leaf extract or a mixed extract of a specific extraction raw material/extraction solvent ratio among extracts of licorice by-products. In this case, the specific extraction raw material/extraction solvent ratio is preferably a ratio of 70 to 200 ml of solvent per 10 g of licorice by-product, more preferably a ratio of 80 to 150 ml of solvent per 10 g of licorice by-product, more preferably 10 g of licorice by-product It is the ratio of 90 to 120 ml of solvent per sugar.

In particular, in the case of antioxidant activity, the fact that the mixed extract of the specific extraction raw material/extraction solvent ratio showed higher activity compared to other extracts despite the same concentration was used together with the licorice licorice and the specific extract raw material/extraction as described above. This means that it exerts a synergistic effect when extracting with solvent ratios. Therefore, in the case of a cosmetic composition for use as an antioxidant (prevention of skin aging) or for a use requiring more antioxidant activity based on this, it is preferable to include a mixed extract of the specific extraction raw material/extraction solvent ratio as the extract.

The cosmetic composition of the present invention may be the licorice by-product extract itself as described above, and may further include a cosmetically acceptable carrier or additive. In this case, the carrier also includes a pharmaceutically acceptable carrier. In addition, the cosmetic composition of the present invention can be formulated in various forms, for example, moisture cream, lotion, nutrient lotion, nutrient cream, massage cream, pack and nutrient essence.

The cosmetic composition of the present invention may further include other functional substances in addition to the licorice by-product extract. In this case, the functional material may be, for example, a material obtained by processing licorice by-products by other methods, or extracts or fermented products of other raw materials other than licorice by-products. In addition, the cosmetic composition of the present invention may include only licorice by-product extract as an extract, and may not include extracts of other raw materials.

The cosmetic composition of the present invention may include, for example, 0.1 to 20% by weight of the licorice by-product extract, but is not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention by these examples.

[Example]

1. Preparation of licorice by-product extract

The leaves and crawfish, which are by-products of domestic licorice, were prepared, and each was freeze-dried and powdered.

Creeping chrysanthemum powder 10g (GS10), leaf powder 10g (GL10), a total of 5 g each of the larvae powder and leaf powder, a total of 10g (GS5+GL5), or 10 g each of the larvae powder and leaf powder, a total of 20g (GS10+GL10) ) was used as an extraction raw material, and each extracted with 100 ml of 80% (v/v) ethanol to obtain an extract.

At this time, the extraction was repeated three times under the conditions of about 24 hours at about 60 ℃.

Each extraction yield is shown in Table 1 below.

Extract name Extraction method Extraction raw material amount (g) Extraction yield (%) GS10 Extract 10 g of larvae powder with 100 ml of 80% EtOH 10 30.9 GL10 Extract 10 g of leaf powder with 100 ml of 80% EtOH 10 23.9 GS5+GL5 A mixture of 5 g of larvae powder and 5 g of leaf powder was extracted with 100 ml of 80% EtOH 10 24.5 GS10+GL10 Extract the mixture of 10 g of larvae powder and 10 g of leaf powder with 100 ml of 80% EtOH 20 21.15

2. Cytotoxicity investigation

RAW 264.7 (KCLB 40071, Korea) macrophages used in the experiment were purchased from the Korean Cell Line Bank (KCLB; Korea) and used. The medium used was DMEM (Dulbecco's Modified Eagle Medium; Lonza, USA) in 10% FBS (Fetal Bovine Serum; Gibco, USA) and 1% Penicillin (100U/ml)/Streptocycin (100㎍/ml)/Amphotericin B ( 0.25 μg/ml) (BioWhittaker, Lonza, USA) was added, and the experiment was performed by culturing in an incubator (Thermo Forma, Series II3111, USA) at 37° C., 5% CO ₂ .

The cell viability of the sample was tested using the Enhanced Cell Viability Assay kit (EZ-Cytox, DoGenBioCo., Ltd.) method. Cells were aliquoted in a 96-well plate at a concentration of 5×10 ⁴ cells/well and cultured for 24 hours, followed by removal of the lung medium. Herein, fresh DMEM (10% FBS, 1% Penicillin (100 U/ml)/Streptocycin (100 μg/ml)/Amphotericin B (0.25 μg/ml)) medium (180 μl) was added, and extracts were treated by concentration (20 After incubation for 24 hours, 10 μl of EZ-Cytox reagent was added to each well and reacted in an incubator at 37° C., 5% CO ₂ for 1 hour. Using a microplate reader (Thermo Scientific Multiskan GO, USA) Absorbance was measured at 450 nm.

As a result, as shown in Figure 1, the extract (GS10) extracted only from the larvae powder alone (GS10), the extract extracted only from the leaf powder (GL10), and the mixed extract (GS5+GL5) extracted by adding 5 g each of the larvae powder and leaf powder were 70% or more at all concentrations. Cell viability was shown. However, in the case of the mixed extract (GS10+GL10), which was extracted by adding 10 g each of C. larvae powder and leaf powder, the cell viability was found to be low compared to this. This result is based on the average value of 3 repeated experiments.

3. Investigation of the anti-inflammatory activity of the extract

The anti-inflammatory activity of each extract was investigated by the nitric oxide (NO) production inhibitory effect test.

RAW 264.7 cells were seeded in a 12-well plate at a concentration of 2×10 ⁵ cell/ml, and then the medium was removed after culturing for 24 hours. Here, a sample diluted to 1 mg/ml in fresh DMEM (10% FBS, 1% Penicillin (100 U/ml)/Streptocycin (100 μg/ml)/Amphotericin B (0.25 μg/ml)) medium was pretreated for 1 hour. After treatment with LPS (10ng/ml), it was cultured in an incubator at 37° C., 5% CO ₂ for 23 hours. After 24 hours of sample treatment, the culture solution was centrifuged, and 100 μl of the cell culture supernatant was mixed with 50 μl of Griess reagent solution A (1% sulfanilamide) + 50 μl of solution B (0.1% N-(1-naphtyl)ethylenediamine dihydrochloride). - After reacting in a well plate for 20 minutes, absorbance was measured at 540 nm using an ELISA reader. Each result was applied to a standard curve (Y=0.0105X+0.0037, R ² =0.9998) prepared using 1M NaNO ₂ (Sodium nitrite) to calculate the amount of NO contained in the supernatant of the cell culture solution.

As a result of the experiment by treating each extract at concentrations of 100, 200 and 400 μg/ml, all extracts (GS10, GL10, GS5+GL5 and GS10+GL10) showed high anti-inflammatory activity at all concentrations tested as shown in FIG. In particular, the extracts extracted with leaf powder alone (GL10) and the mixed extract (GS5+GL5) extracted by adding 5 g each of C. larvae powder and leaf powder showed the highest anti-inflammatory activity. This result is based on the average value of 3 repeated experiments.

4. Investigation of the antioxidant activity of extracts

The antioxidant activity of each extract was investigated by measuring the DPPH radical scavenging activity.

A modified method of Blois MS (1958) was used to measure DPPH radical scavenging activity. 180 μl of 0.1 mM DPPH (2,2-diphenyl-1-picrylhydrazyl) solution was added to each well of a 96-well plate, and 20 μl of a sample obtained by dissolving the extract in second distilled water at each concentration was added and reacted at 37° C. for 30 minutes. Then, absorbance was measured at 517 nm. In this case, ascorbic acid was used as a control. DPPH radical scavenging activity was calculated by the following formula.

DPPH radical scavenging activity (%) = 100 - {(absorbance in the sample added group / absorbance in the no sample group) * 100}

As a result, as shown in FIG. 3, all extracts (GS10, GL10, GS5+GL5 and GS10+GL10) showed high antioxidant activity, and in particular, the mixed extract (GS5+GL5) extracted by adding 5 g each of C. It showed the highest antioxidant activity. The fact that the mixed extract showed higher activity among the experimental groups treated with the same concentration means that when the stems and leaves of licorice are used together, they exert a synergistic effect in antioxidant activity.

5. Investigation of antibacterial activity of extracts

Staphylococcus aureus (KCCM 1335), an aerobic gram-positive strain, Escherichia, an aerobic gram-negative strain. coli (KCTC 2571) and Pseudomonas aeruginosa (KCCM 11802), and Candida albicans (KCCM 11282), a fungus causing candidiasis, were investigated for antibacterial activity (see Table 2).

Each strain was purchased from the Center for Biological Resources (KCTC) or Korea Microorganism Conservation Center (KCCM) and used.

strain strain number badge Escherichia coli KCTC 2571 LB Staphylococcus aureus KCCM 1335 TSA Pseudomonas aeruginosa KCCM 11802 TSA Candida albicans KCCM 11282 YEPD

Take out the stock tube of each strain stored at -80℃ and thaw, inoculate S. aureus and P. aeruginosa in TSA medium, E. coli in LB medium, C. albicans in YEPD medium, and a paper disc on it (10 mm, ADVANTEC, Toyo Roshi Kaisha, Ltd., Japan) was mounted, and 8 mg of the extract (used by dissolving in secondary distilled water) was added to a paper disk, and then incubated at 30° C. or 37° C. for 24 to 48 hours. Then, the diameter of the clear zone formed around the paper disk in the medium was measured. At this time, it means that the larger the size of the generated clear zone, the higher the antibacterial activity. It was used while subcultured at intervals once every 7 days so that the activity of the strain did not fall during the experiment.

As a result, as shown in Table 3, all extracts (GS10, GL10, GS5+GL5, and GS10+GL10) against P. aeruginosa and S. aureus showed high antibacterial activity, and against C. albicans , extracts using larvae powder (GS10, GS5+GL5 and GS10+GL10) showed high antibacterial activity. Antibacterial activity against E. coil was not shown.

target microorganism number sample Clear zone size after 24 hours (mm) Clear zone size after 48 hours (mm) E. coil One GS10 - - 2 GL10 - - 3 GS5+GL5 - - 4 GS10+GL10 - - P. aeruginosa One GS10 17±0 17.5±0 2 GL10 14.5±0.71 14.75±0.35 3 GS5+GL5 15±0 16±0.0 4 GS10+GL10 13.75±0.35 16±0 S. aureus One GS10 20.5±0.71 20±0 2 GL10 15±0 15.5±0.71 3 GS5+GL5 17.5±0.71 17.25±1.06 4 GS10+GL10 16.5±0.71 16.5±0.71 C. albicans One GS10 13.75±0.35 14±0 2 GL10 - - 3 GS5+GL5 13±0 13.5±0 4 GS10+GL10 13.5±0.71 13.5±0.71

6. Investigation of total polyphenol content of extract

To 100 μl of each extract solution having a concentration of 0.25 mg/ml, 2 ml of a 2% Na ₂ CO ₃ solution was added, stirred sufficiently, and then left for 3 minutes. Then, 100 μl of 50% Folin&Ciocalteu's phenol reagents was added, mixed, and reacted for 30 minutes in the absence of light, followed by measurement at 750 nm using a spectrophotometer. The polyphenol content was calculated from a calibration curve (Y=0.0018x-0.0087) using gallic acid as a standard material.

As a result, high total polyphenol content was measured in all extracts (GS10, GL10, GS5+GL5 and GS10+GL10) as shown in FIG. The mixed extract (GS5+GL5) extracted by adding each one showed the highest total polyphenol content.

7. Investigation of Total Flavonoid Content of Extracts

After mixing 0.3 ml of 5% NaNO ₃ solution with 2 ml of each extract solution having a concentration of 0.25 mg/ml, the mixture was reacted at room temperature for 6 minutes. Then, 0.3 ml of 10% Al(NO 3 ) 3 was added, followed by reaction at room temperature for 6 minutes, and then 4 ml of a 10% NaOH solution was added and sufficiently stirred. Then, 10 ml of distilled water was filled, mixed, and reacted for 15 minutes, and then absorbance was measured at 510 nm using a spectrophotometer. Total flavonoids were calculated from a calibration curve (Y=0.0026x-0.0007) using rutinhydrate as a standard material.

As a result, as shown in Figure 5, high total flavonoid content was measured in all extracts (GS10, GL10, GS5+GL5 and GS10+GL10). The mixed extract (GS5 + GL5) extracted by adding 5 g of each leaf powder and the mixed extract (GS10 + GL10) extracted by adding 10 g each of the larvae powder and leaf powder showed high total flavonoid content.

Claims (7)

A cosmetic composition comprising an extract of licorice by-product. The method of claim 1,
The licorice by-product is a leaf of licorice, larvae, or a mixture thereof, the composition. 3. The method of claim 2,
The licorice by-product is licorice leaves and creepers 1: 2 to 2: 1, the composition will be mixed in a weight ratio of 1. The method of claim 1,
Wherein the extract is extracted with water, ethanol, or a mixed solution thereof, the composition. 5. The method of claim 4,
The extract is extracted with 50 to 95% (v / v) ethanol, the composition. 4. The method of claim 3,
The extract will be extracted with 70 to 200㎖ of 50 to 95% (v / v) ethanol per 10g of the licorice by-product, the composition. The method of claim 1,
The cosmetic is a cosmetic for skin antioxidant, antibacterial or anti-inflammatory, the composition.

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